248 results about "Protein purification" patented technology

The invention discloses a glycosyl transferase mutant and an application of the glycosyl transferase mutant in synthesis of rebaudioside. According to the invention, single-point mutation and multi-point mutation are carried out on the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, and a mutant with improved catalytic activity is obtained. The catalytic activity, the substrate specificity and / or the substrate specificity of the glycosyl transferase mutant are / is changed, and the catalytic activity of enzyme to a specific substrate can be remarkably improved by mutation at a specific site. And carrying out induced expression and protein purification on the obtained mutation sequence to obtain the mutant enzyme. The mutant enzyme is used as a catalyst, and UDPG is used as a glycosyl donor, so that the catalytic reaction efficiency of substrates such as stevioside ST, rebaudioside A (RebA) and rebaudioside D (RebD) can be obviously improved. According to the glycosyl transferase UGT76G1 mutant constructed by the research, the catalytic activity of the glycosyl transferase UGT76G1 mutant is improved, and efficient production of rebaudioside M is realized by optimizing a reaction system.

The invention discloses a split intein and application thereof, the split intein comprises two independent peptide fragments: (a) an N-terminal fragment, the amino acid sequence of which is as shown in SEQ ID NO.1; (b) a C-terminal fragment or a variant thereof, the amino acid sequence of the C-terminal fragment is as shown in SEQ ID NO.2, and the amino acid sequence of the variant is as shown in SEQ ID NO.3 or SEQ ID NO.4. The split intein has splicing activity and excellent splicing rate, the application of the split intein can be widened, the number of amino acids of the C-terminal fragment or the variant thereof is small, and the split intein has remarkable advantages in protein purification, solid-phase synthesis and the like.

The invention provides a recombined protein rSp14-3-3 of sarcocystis tenella, an antibody and an ELISA (Enzyme-Linked Immunosorbent Assay) diagnostic kit of the recombined protein rSp14-3-3 of the sarcocystis tenella. The research firstly provides the recombined protein rSp14-3-3 of the sarcocystis tenella. According to the present invention, the recombinant protein rSp14-3-3 is used as a gene fragment, the obtained gene fragment is connected to an expression vector to carry out induced expression and protein purification, the purified recombinant protein rSp14-3-3 is used to coat an ELISA plate, the ELISA detection method for sheep infected with sarcosporidium is established, and the specific monoclonal antibody for the recombinant protein rSp14-3-3 is further provided. All the products can be applied to diagnosis of infection of sarcocystis in samples, water samples, food, environmental samples and the like, and technical support is provided for subsequent immunological diagnosis of sheep sarcocystis by using the protein.

The invention discloses a preparation method of a high-stability manual buffer solution, which comprises the following steps of: introducing two types of slow-release porous particle structures, namely a first type of particles loaded with acidic or alkaline components and a second type of particles loaded with silver nanoparticles, so that continuous slow release and dynamic adjustment of the components in the buffer solution are realized; the pH drift and the aging of a buffer system are effectively delayed; meanwhile, slow release of the silver nanoparticles endows the buffer solution with lasting antibacterial protection, microbial growth is remarkably inhibited, and due to the structural design, the long-term pH stability and storage safety of the buffer solution are remarkably improved, the clarity and physicochemical properties of the buffer solution are guaranteed, and the stability of the buffer solution is improved. The method greatly expands the application range in the fields of high-requirement biological experiments, medical detection, long-term sample storage and the like, the particle addition amount and the component proportion can be flexibly adjusted according to actual requirements, the method adapts to different pH ranges and application scenes, and various experiment requirements of protein purification, long-term sample storage, high-sensitivity analysis and detection are met.

The invention relates to a preparative protein electrophoresis purification system and a protein purification method. The system comprises an automatic gel preparation device and an electrophoresis device, the automatic glue making device comprises a supporting frame, a clamping arm and a glue filling needle, and a linear module is fixedly arranged on the supporting frame to drive the glue filling needle to synchronously move in the horizontal direction and the vertical direction. The electrophoresis device comprises an outer electrophoresis tank, an electrophoresis frame and a cooling device, the electrophoresis frame and the cooling device are arranged in the outer electrophoresis tank, an inner electrophoresis tank is arranged in the electrophoresis frame, rubber plates are detachably arranged on the two sides of the electrophoresis frame, the rubber plates form side plates of the electrophoresis frame, notch grooves are formed in the inner sides of the tops of the rubber plates, and the rubber filling needles are matched with the rubber plates. The protein purification method is a method based on the protein electrophoresis purification system.

The invention discloses an AI-assisted protein purification result analysis method and system. The method comprises the following steps: S1, collecting chromatogram data, electrophoresis image data and mass spectrum peak table data; s2, performing data preprocessing after the chromatogram data, the electrophoresis image data and the mass spectrum peak table data are obtained; s3, carrying out AI identification analysis on the preprocessed data to realize electrophoretic band identification, chromatographic peak identification and mass spectrum deconvolution; s4, performing multi-modal fusion on the chromatography, gel electrophoresis and mass spectrum information sources; and S5, automatically generating a report, and producing a visual chart. The deep learning and multi-modal data fusion technology is introduced, comprehensive automatic analysis of chromatographic data, electrophoresis images and mass spectrum results is achieved, then automatic result analysis of the protein purification process is achieved, an artificial intelligence algorithm is used for recognizing a peak structure and an electrophoresis band, multi-modal data fusion is achieved, and the detection accuracy is improved. And generating a visual analysis report according to a data result.

The invention discloses a construction method and application of a chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain, the construction method comprises the following steps: carrying out gene modification on sweeteners Thaumatin and Brazzein, optimizing preference codons of chlamydomonas reinhardtii to synthesize CrThaumatin and CrBrazzein encoding genes, cloning the encoding genes into an expression vector pGM6, introducing the recombinant expression vector into a chlamydomonas reinhardtii wild-type chlamydomonas sp. Algal strain, and carrying out high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii. And screening by using a paromomycin resistance marker carried by the expression vector to successfully obtain an engineering algal strain. According to the present invention, the engineering strain is subjected to fermentation production to obtain the dry and stable Chlamydomonas reinhardtii powder rich in the target sweet protein, and the protein purification oral test results show that the sweet taste of the Chlamydomonas reinhardtii powder is 3-5 times of the sweet taste of the natural Thaumatin and Brazzein protein;

Provided is a technique for improving the efficiency of adsorption of a protein onto a carrier in an apparatus equipped with an in-line mixer and a hydrocyclone. An adsorption method disclosed herein is performed using an apparatus equipped with an in-line mixer and a hydrocyclone connected to the in-line mixer. The adsorption method is a method in which a protein is adsorbed onto a carrier provided with a ligand capable of binding to the protein. The adsorption method includes: mixing a first slurry containing a carrier with a second slurry containing a protein using an in-line mixer to prepare a mixed slurry; and adsorbing the protein onto the carrier in the mixed slurry to produce a complex between the protein and the carrier. The mixed slurry is prepared so as to contain the carrier in an amount of 10-25 vol% and the protein in an amount of 5-20 mg per 1 ml of the carrier when the whole amount of the mixed slurry is 100 vol%. The mixing time in the inline mixer is 0.6-60 seconds.

The invention relates to the technical field of protein purification, in particular to a purification method of label-free A53T mutant alpha-synuclein. The purification method disclosed by the invention is a purification method which is more efficient and economical and can maintain protein functions, 20mg of protein can be obtained by inducing 1L of bacterial liquid for 4 hours through IPTG, and the method can improve the overall effect and experimental accuracy of preparation of the label-free A53T alpha mutant-synuclein monomer.

The invention discloses preparation and application of grouper enterospora sporowall protein SWP26 and a polyclonal antibody of the grouper enterospora sporowall protein SWP26, and belongs to the field of animal quarantine, the grouper enterospora sporowall protein SWP26 is obtained by amplifying grouper enterospora genes by using a PCR (Polymerase Chain Reaction) method, shearing a target band and transferring the target band into a pET-32a vector to construct a recombinant plasmid, and then transforming escherichia coli BL21 (DE3) for induced expression. A Ni-NTA affinity chromatography method is used for protein purification, animal immunization and polyclonal antibody purification are carried out on the purified SWP26 recombinant protein, western blot detection is carried out on the purified antibody, an obvious signal appears at about 43kDa, and it is proved that the anti-SWP26 polyclonal antibody can be subjected to a specific reaction with the SWP26 purified protein. The invention clones and identifies the high-abundance spore wall protein SWP26 positioned on the surface of the enterosporidium of the grouper for the first time, belongs to a specific protein of the enterosporidium of the grouper, and can be used as a drug target for treating enterocytozoonosis of the grouper.

The instant invention relates to compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, e.g., an anti-IL-1R1 antibody or antigen binding portion thereof, and methods, e.g., cell culture and protein purification methods, for producing such compositions.

The utility model relates to a protein high-flux purification device in the technical field of protein purification, which comprises a liquid receiving box, a deep hole plate is sleeved at the upper end of the liquid receiving box, a plurality of deep grooves are arranged at the upper end of the deep hole plate, water outlet holes communicated with the liquid receiving box are arranged on the inner bottom walls of the deep grooves, and a gasket is inserted in the deep hole plate. A pair of rectangular blocks are fixedly connected to a pair of side ends of the deep hole plate respectively; after purification processing, a spray head enters a deep groove, and water is sprayed out of the spray head under the pressure of a rubber plate to flush the inner wall of the deep groove, so that during use, the cleaning steps are simple and time-saving, personnel operation is facilitated, and after cleaning, clean water is sucked into a cover plate to be stored for next cleaning and use; and the deep groove is sealed through the cover plate, so that the interior of the deep groove is not prone to being polluted by dust, and dustproof protection is conducted on the deep groove.

To provide a new method for efficiently purifying an exogenous protein expressed in a plant.SOLUTION: A method for purifying a foreign protein expressed in a plant, comprising the step of adding an acid to an extract of a tissue of a plant in which a foreign protein is expressed to degrade contaminating proteins derived from the plant.SELECTED DRAWING: None

The invention provides a process of purification of antibody or fusion protein from protein mixture comprising product and process related impurities. The process provides the use of hydroxyapatite chromatography for the separation of low molecular weight impurities and basic variants. In addition, invention further provides a scalable purification process to remove product and process related impurities.

The utility model relates to a bovine serum albumin purification device, and aims to improve the dropping precision of a salt solution in a protein purification process through automatic design and accurate control. The device comprises a pump box, a conveying wheel set, a liquid conveying rubber pipe, a driving motor and a liquid storage tank. The driving motor drives the conveying wheel set to rotate, and the abutting roller in the conveying wheel set revolves around the main shaft sleeve to extrude the liquid feeding rubber pipe, so that the salt solution is pushed to move along the pipeline, and accurate dripping of the liquid is realized. A groove gland is arranged on one side of the pump box and used for pressing the liquid conveying rubber pipe when the conveying wheel set is not started, and liquid leakage is prevented. The arrangement of the adjusting disc allows the abutting roller to be adjusted in the radial direction so as to adapt to liquid feeding rubber pipes of different specifications, and therefore the output amount of the salt solution is accurately controlled. The device is simple in structure and convenient to operate, can effectively improve the purification efficiency and quality of bovine serum albumin, and is suitable for various experiments and production environments.

1. Name of the product in this design: Fully Automated Protein Purification Instrument. 2. Application of this design: For fully automated and efficient purification of various proteins. 3. The key design feature of this product is its shape. 4. The image or photograph that best illustrates the design's key points: a 3D model.

The invention discloses a purification method of a GSL5 protein with disease-resistant and disease-susceptible dual functions, belongs to a protein purification technology, and provides a purification method of a plant gene-GSL5 expression protein with disease-resistant and disease-susceptible dual functions, which comprises the step of fusing two different affinity tags at the C terminal of GSL5, the high-purity target protein GSL5 is obtained through a two-step mild and efficient affinity purification mode, a complex multi-step purification process is avoided, the purification efficiency is improved, and the method can be used for three-dimensional structure research of GSL5 and drug molecule design research based on the structure of GSL5.

The invention discloses a preparation method and application of carassius auratus rahn aquatic OmpA gene cloning and prokaryotic expression as well as a polyclonal antibody. The preparation method comprises the following steps: culture and DNA amplification of rahn aquatic, design of an OmpA primer, gene cloning and prokaryotic expression, back multi-point subcutaneous injection of an immune rabbit, heart blood sampling and preparation of the polyclonal antibody. The invention further provides OmpA protein prokaryotic expression and protein purification, preparation of OmpA polyclonal antiserum and application of fish bodies. According to the invention, a rahnella aquatilis OmpA gene is cloned, a specific Hind III and EcoR1 double-enzyme digestion primer sequence OmpA-1F / OmpA-1R is designed, a prokaryotic expression vector pET32a-OmpA is constructed, high-immunogenicity OmpA protein is obtained through purification, OmpA specific multi-antibody serum is prepared, and the OmpA specific multi-antibody serum can be used for preparing the multi-antibody serum. The OmpA polyclonal antibody can be used for detecting and positioning the expression application of rahnella aquatica OmpA protein in crucian carp bodies through bidirectional agar diffusion, an immunoblotting method and an immunohistochemical method, and provides a reference basis for research and development and application of rahnella aquatica subunit vaccines and a molecular diagnosis technology as well as immune prevention and control of infectious diseases of the rahnella aquatica subunit vaccines and the molecular diagnosis technology.

Provided is a technology for improving the efficiency of separating a complex of a protein to be purified and a carrier in a hydrocyclone of a device including an in-line mixer and the hydrocyclone. A protein purification device disclosed herein comprises a first supply unit, a second supply unit, an in-line mixer, a hydrocyclone, and a dilution line. The first supply unit supplies a first slurry containing a carrier including a ligand that binds to a protein. The second supply unit supplies a second slurry which is a cell culture solution containing a protein. The in-line mixer mixes the first slurry and the second slurry. The hydrocyclone separates, downstream of the in-line mixer, a complex of the protein and the carrier from a mixed slurry of the first slurry and the second slurry. The dilution line is provided between the in-line mixer and the hydrocyclone and introduces a diluted liquid into the mixed slurry.

The invention relates to a magnetic bead mediated protein purification device and a high-purity extraction method, and belongs to the technical field of protein purification. The magnetic bead mediated protein purification device comprises a bottom seat, the upper end of the bottom seat is fixedly connected with a circular shell, and the inner bottom of the circular shell is fixedly connected with a bottom magnet; the purification mechanism is mounted on the inner bottom wall of the circular shell; wherein the purification mechanism comprises a plurality of magnetization assemblies mounted on the inner wall of the bottom seat; according to the magnetic bead mediated protein purification device and the high-purity extraction method, the device drives the liquid and the magnetic beads to be mixed through the arranged magnetizing assembly, and after mixing is completed, relatively uniform magnetic attraction can be formed on the liquid storage pipe through the magnetizing assembly, so that the magnetic beads can be uniformly distributed on the inner wall of the liquid storage pipe, and the magnetic beads are uniformly separated from the liquid storage pipe; the occurrence of magnetic bead breakage is reduced, and the extraction effect of protein purification is improved.

1. The name of the design product: protein purification instrument. 2. The use of the design product: biological reaction instrument for protein chromatography separation. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view 1.

The invention belongs to the technical field of protein purification, and particularly relates to a chromatography medium, a preparation method of the chromatography medium and application of the chromatography medium to extraction of alpha-lactalbumin. The chromatography medium provided by the invention takes agarose as a matrix, and the surface of the chromatography medium is modified by a ligand through a glucan spacer arm; wherein the molecular weight of the glucan is 20 kDa to 500 kDa, the ligand comprises propylene diamine, and the density of the ligand in the chromatography medium is 200 [mu] mol / mL to 300 [mu] mol / mL. The invention also provides a preparation method of the material, and the material can be obtained by sequentially carrying out epoxy activation, glucan modification, secondary activation and ligand bonding on the matrix. When the chromatography medium provided by the invention is used for extracting alpha-lactalbumin from whey, the chromatography medium has the advantages of high loading capacity, high loading flow rate and efficient regeneration, and meets the requirements of industrial production.

The invention belongs to the technical field of action target screening and identification, and particularly relates to a method for screening and identifying a Nav channel protein binding agent based on ligand-induced protein stability change. The method comprises the following steps: firstly, carrying out mild lysis on cells of high-expression voltage-gated sodium ion channel protein (Nav channel protein) to obtain lysate containing active Nav channel protein, carrying out contrast incubation on the lysate, a compound to be detected and a solvent, carrying out restrictive enzymolysis by using protease in the presence of a surfactant, and carrying out freeze-drying to obtain the high-expression voltage-gated sodium ion channel protein (Nav channel protein). Representing the retention amount of the target protein and the internal reference protein, and evaluating the protective capability of the compound to be detected on degradation of the target protein to judge the affinity. The method has the advantages of high identification speed, economy and high efficiency, overcomes the problems that the existing Nav channel protein binding agent screening method is tedious, high in technical difficulty and depends on special large instruments, and can be used for screening action targets without specially constructing cells and purifying proteins by large instruments, so that the screening efficiency is effectively improved, and the screening cost is effectively reduced.

The invention provides a full-automatic detection device and method for the concentration of free drugs in blood. The full-automatic detection method adopts a full-automatic detection device and comprises the following steps: (1) loading a blood sample to a protein purification chromatographic column by adopting a pump 1 and an automatic sampler, removing protein components in the blood sample, and retaining free drug components on the protein purification chromatographic column; (2) enabling free drug components in the protein purification chromatographic column to flow into a trapping column and enriching the free drug components in the trapping column; and (3) enabling the free drug components in the trapping column to flow into an analytical column, separating, and detecting in a triple quadrupole mass spectrometer. Through the specific full-automatic detection device, the problems that total drug concentration monitoring cannot meet clinical treatment requirements, and the free drug pretreatment process is complex, long in time and high in cost are solved, clinical individualized drug administration can be assisted, the drug curative effect is improved, and the toxic and side effects of drugs are reduced.

The current invention relates to an improved microfluidic device for protein / protein complex purification, as well as an associated methodology and system.In particular it relates to systems and methods for electron microscopy, preferably cryogenic electron microscopy (cryo-EM), on protein samples, comprising: a. at least one microfluidic chip with inlet and outlet and purification device; b. at least one illumination means and detection means for, preferably fluorescence, measurements on the microfluidic chip; c. a pumping system adapted for operable connection to the microfluidic chip, and configured for controlling flow in the operably connected microfluidic chip; d. a microscopy grid, preferably a cryogenic electron microscopy (cryo-EM) grid, for holding fluid samples; e. preferably a cryogenic container, for a cryogenic coolant; f. preferably a transport system for moving the cryo-EM grid between a position for receiving a fluid sample from the microfluidic chip and the cryogenic container; g. a control system, preferably a processor, which receives information on the measurements from the detection means, configured for controlling the pumping system at least based on said information, and further configured for controlling the illumination and detection means and preferably also for controlling the transport system.

The utility model relates to the technical field of protein purification, and particularly discloses a portable low-temperature protein purification box which comprises a box body, a control panel is mounted at the top of the box body, a temperature sensor is mounted on the inner wall of the box body, and a sample loading module, a purification module, a detection module and a sample collection module are arranged in the box body. The sample loading module comprises a sample liquid storage device, a buffer liquid storage device and a peristaltic pump which are mounted in the box body, the purification module comprises a chromatographic column connected with the peristaltic pump, and the detection module comprises an ultraviolet detector mounted in the box body. Through the design of the low-temperature control system and the heat preservation layer, protein purification can be carried out in a low-temperature environment, the activity of the protein is effectively maintained, and the problem that the protein is inactivated or even degraded due to the fact that the temperature is too high in a traditional purification method is solved; meanwhile, the influence of long-time low-temperature experiments on the body health of experiment operators is avoided.

In situ Raman spectroscopy methods and systems are provided for the characterization or quantification of a protein purification intermediate and / or the final concentrated fraction during production or preparation. In one embodiment, in situ Raman spectroscopy is used to characterize or quantify critical quality attributes of protein purification intermediates during downstream processing (i.e., after the protein purification intermediate has been obtained). For example, the described in situ Raman spectroscopy methods and systems can be used to characterize and quantify protein purification intermediates as the protein purification intermediates are purified, condensed, or otherwise formulated into the final pharmaceutical product to be marketed or administered.

The application belongs to the technical field of biology, and particularly relates to a SUMO protein, a specific protease thereof and application in protein purification. The amino acid sequences of the SUMO specific protease and the SUMO protein are shown in SEQ ID NO. 1 and SEQ ID NO. 3 respectively. The SUMO protein provided by the application can significantly improve the soluble expression amount of a target protein. More importantly, compared with a commercial scSUMO / Ulp1 system, the SUMO specific protease of the application exhibits higher biochemical robustness in a complex buffer, and can maintain high efficient specific cleavage capacity in a wide temperature range and in a high concentration sodium ion and high concentration imidazole environment. The application overcomes the bottleneck of traditional purification process, i.e. complicated dialysis desalting, and can directly perform in-situ efficient enzymatic cutting on a chromatography eluent, and has great application potential and economic value in industrialized production of recombinant protein purification.

The invention relates to the field of protein purification. The present invention envisages a method for purifying a fermentation-derived solution of a protein of interest, the method comprising the steps of: contacting the fermentation-derived solution with chemically activated carbon in an amount ranging from 0.1 wt% to 15 wt% of the solution; adjusting the pH to a value of 4 to 12, and adjusting the temperature to a value of 2 DEG C to 60 DEG C; separating the chemically activated carbon from the solution; and thereby obtaining a clarified solution of the protein of interest. Furthermore, the invention relates to a clarified solution of a protein of interest obtainable or obtained by the method according to the invention, and the use of chemically activated carbon for purifying a fermentation-derived solution of a protein of interest.

The invention relates to a preparation method of a babesiella bovis vaccine, belongs to the technical field of vaccines, and particularly relates to a combined vaccine based on silver nanoparticles (AgNPs) and theileria bovis antigen, the vaccine comprises 10-50nm AgNPs, the surface of the AgNPs is modified with carboxyl, amino or PEG, and the AgNPs are used for covalently coupling RAP-1 protein or immunogenic fragments thereof and MSA-2 protein or B cell epitope peptide thereof, the RAP-1 protein comprises an amino acid sequence (nucleotide sequence is as shown in SEQ ID NO: 1), the MSA-2 protein comprises an amino acid sequence (nucleotide sequence is as shown in SEQ ID NO: 2), and the preparation method comprises the following steps: carrier construction, recombinant expression, protein purification and AgNPs coupling to form a silver nanoparticle antigen compound, and the silver nanoparticle antigen compound is coated with chitosan to prolong the in-vivo half-life period.